Re: Removal of DAB Chromogen

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From:"J. A. Kiernan" <> (by way of histonet)
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At 05:18 PM 12/14/99 -0500, Richard Cartun wrote:
>We had a problem in the lab today where our cytokeratin cocktail
>(AE1&AE3/CAM5.2) was put on sections of prostate instead of the requested
>cytokeratin 34BetaE12 mAb.  Is there any way to remove the DAB reaction
>product and re-stain the slides using the correct mAb?  If we re-stain the
>slides do I have to be concerned with binding of the detection reagents to
>the first primary mAb?

  Nothing will bemove the brown DAB product. You could rinse the
  sections in acid (e.g. about pH 2) to remove all bound antibodies,
  then immunostain again for the correct antigen, using a differently
  coloured detection system, such as AEC for peroxidase (red) or
  an alkaline phosphatase-based ABC or APAP kit (commonly purple
  or red). Nickel- or cobalt-enhanced DAB (blue-black) might be OK.

                    John A. Kiernan
                    Dept of Anatomy and Cell Biology
                    University of Western Ontario
                    LONDON,   Canada.


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